不同程度的过劳对大鼠动脉血管平滑肌细胞的影响  

Effects of different levels of overworking on rat arterial smooth muscle cells

作　　者：甘露 陈苏衡 庄苗 张小晓 郭鸿 黄蓉蓉 李玉兰[2] GAN Lu;CHEN Suheng;ZHUANG Miao;ZHANG Xiaoxiao;GUO Hong;HUANG Rongrong;LI Yulan(The First Clinical Medical College,Lanzhou University,Lanzhou,Gansu Province,730000;Department of Anesthesiology,the First Hospital of Lanzhou University,Lanzhou,Gansu Province,730000,China)

机构地区：[1]兰州大学第一临床医学院,兰州730000 [2]兰州大学第一医院麻醉科,兰州730000

出　　处：《陆军军医大学学报》2022年第5期461-468,共8页Journal of Army Medical University

基　　金：甘肃省卫生健康行业科研项目(GSWSKY2021-079)。

摘　　要：目的研究不同程度的过劳对大鼠动脉血管中层平滑肌细胞(vascular smooth muscle cells,VSMC)的影响。方法将清洁级SD大鼠分为对照组(CON)、过劳组(OW)及过劳+睡眠不足组(OW+SD)。OW组采用力竭游泳法建立过劳模型;OW+SD组在力竭游泳的基础上加水中平台法限制睡眠时间。15 d后取出大鼠腹主动脉,行HE染色进行病理学观察并测量中膜厚度,透射电镜观察细胞内超微结构,用TUNEL法检测各组大鼠腹主动脉中层VSMC的凋亡率,用RT-qPCR法检测VSMC中Caspsae-3、Bax、Bcl-2和Ki67的mRNA表达水平,用免疫组化法检测血管中层凋亡蛋白Caspase-3的含量。结果(1)HE染色示:与CON组相比,OW组和OW+SD组大鼠血管壁VSMC排列紊乱,有"空泡"样细胞形成,血管中膜厚度较CON组明显增加(P<0.05);(2)电镜示:OW组和OW+SD组大鼠VSMC细胞中可见凋亡小体形成;(3)TUNEL结果显示:OW组和OW+SD组大鼠VSMC凋亡率明显高于CON组(P<0.05);(4)RT-qPCR结果显示:与CON组相比,OW组和OW+SD组血管中膜Bax、Ki67的mRNA表达明显增加(P<0.05),OW组和OW+SD组血管中膜Caspase-3和Bcl-2的mRNA表达无明显变化(P>0.05);(5)免疫组化结果显示:与CON组相比,OW组和OW+SD组血管中膜Caspase-3的蛋白含量明显增加(P<0.05)。结论过劳可引起大鼠动脉血管中层VSMC凋亡,诱发动脉血管壁中层VSMC异常增殖和修复,可形成动脉病变的早期病理基础;并且在实验时间内,睡眠未能改善过劳对血管平滑肌细胞的损伤。Objective To determine the effect of overworking on vascular smooth muscle cells(VSMC)in rats.Methods Clean SD rats were randomly divided into control group(CON),overworking group(OW)and overworking and sleep deficiency group(OW+SD).The rat model of overworking was established by exhaustive swimming in the latter 2 groups,and the rats of the last group was subjected to sleep deprivation with an underwater platform.In 15 d later,the abdominal aorta of rats was collected for pathological observation by HE staining,and the thickness of media was measured with aid of Image J software.The intracellular ultrastructure was observed by transmission electron microscopy.The apoptotic rate of VSMC was detected by TUNEL,the mRNA expression of Caspsae-3,Bax,Bcl-2 and Ki67 in the VSMC were detected by RT-qPCR,and the content of apoptotic protein Caspase-3 was measured by immunohistochemical assay.Results(1)The results of HE staining showed that VSMC were in disordered arrangment and formed vacuole-like cells in the OW and OW+SD groups,and the vascular intima-media thickness was significantly thicker in the OW and OW+SD groups than the CON group(P<0.05).(2)Electron microscopy displayed the apoptotic bodies in the VSMC of the OW and OW+SD groups.(3)TUNEL results showed that the apoptotic rate of VSMC was significantly higher in the OW and OW+SD groups than the CON group(P<0.05).(4)RT-qPCR showed that the expression levels of Bax and Ki67 were significantly increased(P<0.05),while those of Caspase-3 and Bcl-2 were not obviously changed in OW and OW+SD groups(P>0.05)when compared with the CON group.(5)Immunohistochemical results showed that the protein content of Caspase-3 in vascular middle membrane of the OW and OW+SD groups was increased significantly than that in the CON group(P<0.05).Conclusion Overworking aggravates the apoptosis of VSMC and induces the proliferation and repair of VSMC,which form the pathological basis of early arterial lesions.During the experimental time,sleep could not alleviate the injury of VSMC caused

关 键 词：过劳 动脉血管壁 血管平滑肌细胞 凋亡 增殖 

分 类 号：R163[医药卫生—公共卫生与预防医学] R322.12